The Vector Core will design, construct, produce and characterize adeno-associated virus in a facility equipped specifically for the high through-put production of viral vectors and provide multiple preparations of these vectors to the Program Projects. The Vector Core staffhas extensive experience, including participation as the national service core of the NHLBI Programs in Excellence in Gene Therapy and has developed a comprehensive set of reagents and protocols for gene transfer and expertise in their use. To support this Program Project the Core will closely coordinate the schedule of vector production with the Projects PI's to provide AAV vectors of numerous serotypes, reporter genes, segmental and full cDNA versions of the CFTR gene, under the control of promoters as relevant for the aims. Continuous vector production and on-going methods development for the new vector constructs is a task that best fits the core model in that significant expertise is required and the overall strategy across vector types is similar. The quality of the laboratory grade vectors for in vitro and in vivo applications of 3 of the projects will be verified with analytical methods developed at the Vector Core using standards established for preclinical development in the GMP facility at the Belfer Gene Therapy Core Facility. This quality control mechanism is an integrated part of the Vector Core's program to support the projects and minimize artifactual results. For each of the AAV serotypes the Core uses procedures developed for AAV production that have been optimized for yield, purity and stability of product. Further refinement of methods for each serotype will be provided to assure the quality of vectors prior to use by each of the program projects. The Core estimates it will produce 42 vectors each year for the projects.